JOURNAL ARTICLE
RESEARCH SUPPORT, NON-U.S. GOV'T
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Distribution of NADPH-diaphorase and expression of nNOS, N-methyl-D-aspartate receptor (NMDAR1) and non-NMDA glutamate receptor (GlutR2) genes in the neurons of the hippocampus after domoic acid-induced lesions in adult rats.

Neuronal degeneration followed by detection of nitric oxide (NO)-producing neurons of the hippocampus was investigated at 4 h, 16 h, 24 h, 2 days, 5 days, and 14 days after administration of domoic acid (DA), in the present study. Histopathological analysis (Nissl staining) displayed dark-stained degenerating neurons in the hippocampus at 24 h to 14 days after DA administration, with degeneration most severe at 5-14 days. NADPH-d-positive neurons were observed in different subfields of the hippocampus in control rats and DA treated rats at 4-24 h. Complete loss of NADPH-d-positive neurons in the CA1 and CA3 subfields and also in the hilus of dentate gyrus (DG) was observed at 5 days and 14 days after the administration of DA. In contrast, at 4-24 h, neuronal nitric oxide synthase (nNOS)-immunoreactive cells were absent from the hippocampal subfields in control and DA-treated animals but were observed at 5 days and 14 days after DA administration. N-methyl-D-aspartate receptor (NMDAR1) immunoreactivity was increased in the hippocampal neurons at 5 days after DA administration and double immunofluorescence demonstrated its coexpression with induced nNOS expression. No significant change could be observed in the immunoreactivity of non-NMDA receptor (GlutR2) as compared with the controls, while occasional immunoreactive neurons were colocalized with induced nNOS expression. Reverse transcription-polymerase chain reaction analysis showed the upregulated expression of nNOS and downregulated expression of NMDAR1 at 5 days after the administration of DA. Although nNOS mRNA expression was rapidly induced at 5 days after DA administration, in situ hybridization analysis revealed complete loss of nNOS mRNA expression in the region of neuronal degeneration in the hippocampus at 24 h and 5 days after DA administration. The present study has shown that NADPH-d and nNOS express differentially in the neurons of the hippocampus in DA-induced neurotoxicity. It is speculated that induction of nNOS and glutamate receptor genes in the neurons of the hippocampus in response to DA-induced neurotoxicity could have contributed to the neuronal degeneration.

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